The bioactivity of a working reference standard was determined by replicate bioassays with calibration against a primary reference standard. In this study the number of bioassay replicates needed for calibration first was calculated theoretically, and if the mean value of the experimental bioassay replicates fell within the predefined bioactivity level the bioactivity of the working reference was defined as 100%. Our results showed that when the total intermediate precision of the bioassay method was at 11.66% and the predefined bioactivity level was set at 95%-105% with a confidence level of 95%, 21 bioassay replicates should be carried out for calibration. The average value of the 22 experimental bioassay replicates was 101.96%, so the bioactivity of the working reference standard was consistent with that of the primary reference standard at 100%. The results suggest that a strategy of first calculating the number of bioassay replicates needed for calibration and then determining whether the resulting experimental mean value is within the predefined bioactivity level will be of value to the biopharmaceutical industry.

"Prescription embodied in Preparation", Chinese medicine preparation, aims to study the specific form of Chinese medicine from raw materials to preparation for acting on patients directly. Its development has gone through three stages according to the characteristics of raw materials pretreatment, including "direct smash and initial extraction for Chinese materia medica", "Extensive extraction and preliminarily impurity for Chinese materia medica" and "Refining and purification for Chinese materia medica". With the development of new technologies and new theories, Chinese medicine preparation emerged in a new stage: structural components of Chinese medicine, with the characteristics of definited material basis, clear mechanisms, determined ADME/T properties, reasonable drug release system designs and scientific productions quality controls. This requires multidisciplinary to solve systemly the problems of Chinese medicine preparation. In this article, we reviewed the development of Chinese medicine preparation in different times, and analyzed the development and the characteristics of Chinese medicine preparation; and mainly focused on a fact that multidisciplinary promoted the study and development of Chinese medicine preparation, especially in structural components of Chinese medicine. It provides development direction and theoretical basis for Chinese medicine preparation.
Materia Medica ; isolation & purification ; Medicine, Chinese Traditional ; Technology, Pharmaceutical

Background: Creatine transporter (CRTR) deficiency is the most common creatine deficiency syndrome, of which the final diagnosis relies on mutation in the X-linked CRTR gene. To date, more than 90 mutations in the SLC6A8 gene have been reported. This paper discusses a novel mutation detected via the thorough sequencing of all the X-chromosome-specific exons investigated in a four and a half year old boy with an intellectual disability, speech and language delay and motor disturbance. Methods: A brain magnetic resonance imaging (MRI) and a proton magnetic resonance spectroscopy (MRS) were carried out, the creatine and creatinine concentrations in the urine were checked and all exons were sequenced. Results: A detailed clinical investigation revealed a reduction in the cerebral creatine levels in the brain by the MRS, elevated creatine and creatinine concentrations in the urine and signal abnormalities in the left frontal cortex of the brain by the MRI. A novel change was identified in the heterozygosity of the exon 10: c.1395-c.1401 deletion. Conclusion: The use of a combination of powerful new technologies, such as thorough exome-nextgeneration sequencing and a brain MRS, should be considered, in order to determine any neurometabolic diseases, especially when the signal abnormalities in the brain MRI cannot be explained by any other factors. This mutation results most likely in a dysfunction of the creatine transport and synthesis, hence causing central nervous system symptoms.
Carrier Proteins

Objective To detect the histological characteristics of collagen nodules from hypertrophic scars (HS) and investigate the origin of collagen nodules.Methods The scar tissues were collected from patients with plastic operation.Morphological characteristics of collagen nodules were observed by light microscopy of HE-stained sections; expressions of type Ⅰ /Ⅲ collagens were observed by polarized light microscopy of sirius red-stained sections; expression and distribution of myofibroblasts (MFb)-specific protein (α-smooth muscle actin,α-SMA) were observed by immunostaining in order to observe level of local tissue tension.Results Collagen nodules varied in shape,not only sphereshaped,and in size.Moreover,abundant fibroblasts (Fb) with large and light-stained nuclei were seen in the nodules compared to non-nodule area,indicating that the cells located in the modules were active.Some collagen nodules were composed largely of collagen type Ⅲ (green),but some mainly contained collagen type Ⅰ (red or yellow),indicating the difference in the time of nodule formation.α-SMA was expressed mainly in the deep dermis equivalent to the level of reticular layer of the new scar tissues (2 months after injury) ; α-SMA was expressed mainly in the nodules of the old scar tissues (2-10 years after injury),but almost not in non-nodule areas except for a strongly positive staining in the vessels.Moreover,α-SMA presented a heterogeneous distribution in the collagen nodules,with stronger expression in the epidermal end than in the subcutaneous tissue end and stronger expression in the superficial dermis than that in the deeper part.It was suggested that there existed massive amount of MFb and high tension in the nodules arid that the tension distribution was not uniform in or between the nodules.Conclusions Collagen nodules are of varying shape and size and collagen types are associated with the time of nodule formation.Moreover,Formation of the collagen nodules may be closely related to the distribution and evolution of the local tissue tension.

Objective To evaluate the effects of regular follow-up and systematic management in premature infants on early discovering of cerebral palsy and intervention efficacy in early phase. Methods Fifty premature infants with cerebral palsy recruited from early children development outpatient service, were classified as treatment group; 40 premature infants with cerebral palsy recruited from general outpatient service, were classified as control group. The infants of treatment group received systematic follow-up and intervention. while the infants of control group received no systematic management. Age and therapeutic efficacy were compared between the two groups. ResultsThe average age in the treatment group ( [4. 0 ± 1.2] months) was significantly younger than that of the control group ( [7.0 ± 1.4] months) ( P ＜ 0. 05 ). The mental development index ( MDI )and physical development index (PDI) in the treatment group was 91.8 ± 10. 2 and 90. 2 ± 11.2,respectively,which were significantly higher than that of the control group(80. 2 ± 11.7 for MDI and 79. 3 ± 10. 2 for PDI)( P ＜ 0. 05 ) . Conclusion Systematic management could help discover cerebral palsy in premature infants in time and could improve physical and mental development of these infants.

To investigate the prevalence of Enterococcus faecalis in saliva and filled root canals of patients requiring endodontic retreatment for apical periodontitis. Patients with apical periodontitis who were referred for endodontic retreatment were examined. The type and quality of the restoration, symptoms, quality of obturation were recorded. During retreatment, an oral rinse sample and root canal sample were cultured using brain-heart infusion agar and bile esculinazide agar to select for E. faecalis. The 16S rRNA technique was used to identify E. faecalis. A total of 32 women and 22 men (mean age: 38 years; s.d.: 11 years) and 58 teeth were studied. The prevalence of E. faecalis was 19% in the saliva and 38% in the root canals. The odds that root canals harbored E. faecalis were increased if the saliva habored this bacterium (odds ratio=9.7; 95% confidence interval=1.8-51.6; P<0.05). Teeth with unsatisfactory root obturation had more cultivable bacterial species in root canals than teeth with satisfactory root obturation (P<0.05). E. faecalis is more common in root canals of teeth with apical periodontitis than in saliva. The prevalence of E. faecalis in root canals is associated with the presence of E. faecalis in saliva.
Adolescent ; Adult ; Aged ; Chi-Square Distribution ; Colony Count, Microbial ; Dental Pulp Cavity ; microbiology ; Dental Restoration Failure ; Enterococcus faecalis ; isolation & purification ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Multivariate Analysis ; Periapical Periodontitis ; microbiology ; therapy ; Quality of Health Care ; RNA, Ribosomal, 16S ; genetics ; Retreatment ; Root Canal Obturation ; Saliva ; microbiology ; Young Adult

OBJECTIVETo observe clinical efficacy in treating cutaneous deficiency of traumatic shank amputation stump with full-thickness skin graft combined with vacuum sealing drainage.

METHODSFrom September 2009 to December 2012, 15 patients with cutaneous deficiency of traumatic shank amputation stump were treated with full-thickness skin graft combined with vacuum sealing drainage. Among patients, there were 11 males and 4 females with an average age of 41.5 (ranged from 25 to 62) years old. Ten cases were caused by traffic accident and 5 cases were caused by heavy object, 9 cases on left and 6 cases on right. Six patients with smashed wound were treated with debridement and amputation, combined with vacuum aspiration in-emergency; 9 patients caused by infection and necrosis were treated with debridement and amputation, combined with vacuum aspiration, and full-thickness skin graft were performed at stage II. The skin defect area of residual limbs ranged from 40 cm x 20 cm to 25 cm x 15 cm.

RESULTSAll patients were followed up from 3 months to 1 year. Full-thickness skin graft of residual limbs were survived,and obtained satisfactory walking function with prosthetic. Residual skin increased thicken, wearproof without rupture and pain.

CONCLUSIONFull-thickness skin graft combined with vacuum sealing drainage in treating cutaneous deficiency of traumatic shank amputation stump could reserve the length of residual limbs, increase survival rate of skin graft with less scar of survival skin, get good wearability and it is conducive to prosthetic wear. It is a simple and easy treatment method.

Adult ; Amputation Stumps ; surgery ; Female ; Humans ; Leg Injuries ; surgery ; Male ; Middle Aged ; Negative-Pressure Wound Therapy ; methods ; Skin ; injuries ; Skin Transplantation

AIM: To investigate the clinical effect for treating bullous keratopathy ( BK ) by anterior corneal stromal acupuncture combined with amniotic membrane transplantation.
METHODS: Totally 35 patients ( 35 eyes ) with bullous keratopathy were treated by corneal stromal acupuncture combined with amniotic membrane transplantation. All patients preoperative and postoperative underwent anterior segment OCT, corneal topography, corneal sensitivity and confocal microscopy. To observe postoperative ocular symptoms in patients with BK, recurrence of bulla, changes in corneal thickness, corneal sensitivity and changes in the organizational structure of the layers of the cornea.
RESULTS: Thirty-five were followed up for 6-18mo. The symptom of pain disappeared in 32 cases ( 91%) in the first day after operation and did not recur during follow-up. The symptom of pain relieved in 3 cases (9%) in the first day after operation and disappeared in 3 days. Corneal epithelium of 9 cases ( 26%) were all healed within 1wk, 21 cases (60%) were all healed within 2wk, and 5 cases (14%) were all healed within 3wk. Following up for 6-18mo, there was no recurrence of symptoms or bulla. A small amount of tiny bubbles in the surrounding area appeared in 2 cases after 3 and 4wk. All patients had no neovascularization, and had smooth corneal surface. The amnions of 30 cases (86%) were thinning after 2mo, partially dissolved and absorped, not seen with the naked eye after 3mo. Thirty-four cases (97%) had no changes in vision, one case (3%) was from the light to front of the manual. After 2mo, corneal sensation decreased in 30 cases ( 86%) , corneal thickness increased from preoperative 788±35μm to 940±43μm. After 12mo, corneal thickness increased to 1060±27μm. Results of confocal microscopy: after 3mo, the number density of the trigeminal nerve fibers under corneal basement membrane reduced, shallow stromal cells became into fibrotic stroma, deep stroma was more loose, and cells swelled significantly. The number of endothelial cells reduced and form swelled more obviously compared with preoperatively.
CONCLUSION: Corneal acupuncture combined with amniotic membrane transplantation can effectively control the symptoms of BK, prevent the recurrence of BK, and especially it is a simple, safe and practical way for patients with poor visual function.

The 1.23 kb DNA fragment encoding the early protein EP0 of pseudorabies virus (PRV) Ea strain was amplified by PCR technique and cloned into pBluescriptII sk+.Three sequencing plasmids containing the partial fragment of the EP0 gene were constructed and the sequences were obtained by Sanger's sequencing technique. Compared with PRV InFh strain, there were multipile site-mutations and a deleted-mutation in the EP0 gene of PRV strain Ea，and the diversity of amino acid residues also existed.Then, the EP0 gene was inserted into an expression vector, pET-28a, fused into the downstream of the 6ΧHis-Tag in frame, to yield the expression plasmid pETEP0. After induction by IPTG, a high expression of fusion protein was obtained, SDS-PAGE analysis and Western blotting showed that the fusion protein was 62kD and the protein was specific to antisera against PRV Ea strain. This indicated that the EP0 gene be expressed in BL21(DE3) and the expression products have immuno-genicity.

OBJECTIVETo investigate the effect of augmentative locking compression plate combined with bone graft in treating aseptic femoral shaft nonunion after intramedullary nailing.

METHODSTwenty-one cases with aseptic femoral shaft nonunion after intramedullary nailing from January 2007 to January 2013 were treated,including 18 males and 3 females with a mean age of 37.7 years (ranged from 23 to 64 years). The mean period of nonunion after surgery was 23.9 months (ranged from 9 to 62 months). According to Weber-Cech classification,10 of those 21 cases were hypertrophic nonunion,7 were atrophic, and 4 had oligotrophic fracture nonunion. All patients retained the original intramedullary nail, and applied with augmentation plating of 6 to 8 holes locking compression plate, unicortical fixation with 2 to 3 locking screws in the proximal or distal end, with simultaneous autologous iliac bone grafting. After treatment,all patients were allowed to partial weight-bearing until full weight-bearing according to the radiological results. All patients were followed up and were evaluated with clinical and imaging results.

RESULTSAll patients were followed up from 8 to 24 months, averaged (13.5±3.5) months,which showed clinical union at 4 to 8 months, averaged (6.0±1.0) months and radiological solid union at 7 to 12 months, averaged (9.1±1.5) months. No such complications as infection,hardware loosening or breaking were found.

CONCLUSIONAugmentative locking compression plate(LCP) combined with bone graft for aseptic femoral shaft nonunion after intramedullary nail has a satisfied clinical efficacy. It's an useful and simple method.

Adult ; Bone Nails ; adverse effects ; Bone Plates ; Bone Transplantation ; Female ; Femoral Fractures ; complications ; surgery ; Follow-Up Studies ; Fracture Fixation, Intramedullary ; adverse effects ; Fractures, Ununited ; complications ; surgery ; Humans ; Male ; Middle Aged ; Postoperative Complications ; etiology ; surgery ; Treatment Outcome ; Young Adult